ABSTRACT Craniosynostosis is a debilitating pediatric condition characterized by premature cranial suture fusion, resulting in abnormal skull shape, blindness and neurologic deficits. The prevalence of craniosynostosis is at approximately 1 in 2,500 live births, which is one of the highest incidences of congenital malformations of the skeletal system. We have assembled a strong collaborative team combining Dr. Ma's expertise in 3D tissue- engineering scaffolds and biomolecule delivery for craniofacial tissue regeneration, Dr. Mishina's expertise in suture development and midline craniosynostosis model, and Dr. Hatch's expertise in the mechanisms underlying coronal craniosynostosis. In the Specific Aim 1, we will develop 3D scaffold-based microenvironment to maintain suture mesenchymal stem cells; in Aim 2, we will establish 3D culture system to regenerate and maintain bone-suture-bone tissue composite; in Aim 3, we will establish the efficacy of engineered scaffolds and controlled release systems for preventing craniosynostosis in vivo. By accomplishing these specific aims, we will develop a novel strategy to cure or diminish the severity and recurrence of craniosynostosis in humans, hence significantly improving patient quality of life.